Live cell microscopy analysis of radiation-induced DNA double-strand break motion

We studied the spatiotemporal organization of DNA damage processing by live cell microscopy analysis in human cells. In unirradiated U2OS osteosarcoma and HeLa cancer cells, a fast confined and Brownian-like motion of DNA repair protein foci was observed, which was not altered by radiation. By analyzing the motional activity of GFP-53BP1 foci in live cells up to 12-h after irradiation, we detected an additional slower mobility of damaged chromatin sites showing a mean square displacement of ≈0.6 μm2/h after exposure to densely- or sparsely-ionizing radiation, most likely driven by normal diffusion of chromatin. Only occasionally, larger translational motion connected to morphological changes of the whole nucleus could be observed. In addition, there was no general tendency to form repair clusters in the irradiated cells. We conclude that long-range displacements of damaged chromatin domains do not generally occur during DNA double-strand break repair after introduction of multiple damaged sites by charged particles. The occasional and in part transient appearance of cluster formation of radiation-induced foci may represent a higher mobility of chromatin along the ion trajectory. These observations support the hypothesis that spatial proximity of DNA breaks is required for the formation of radiation-induced chromosomal exchanges.

[1]  J. Ward,et al.  The complexity of DNA damage: relevance to biological consequences. , 1994, International journal of radiation biology.

[2]  D T Goodhead,et al.  Initial events in the cellular effects of ionizing radiations: clustered damage in DNA. , 1994, International journal of radiation biology.

[3]  Tom Misteli,et al.  Spatial genome organization in the formation of chromosomal translocations. , 2007, Seminars in cancer biology.

[4]  Mariel Vazquez,et al.  Radiation-induced chromosome aberrations: insights gained from biophysical modeling. , 2002, BioEssays : news and reviews in molecular, cellular and developmental biology.

[5]  G. Taucher‐Scholz,et al.  Ion Irradiation as a Tool to Reveal the Spatiotemporal Dynamics of DNA Damage Response Processes , 2006 .

[6]  G. Kraft,et al.  Calculations of heavy-ion track structure. , 1995, Radiation and environmental biophysics.

[7]  G. Taucher‐Scholz,et al.  Positional Stability of Damaged Chromatin Domains along Radiation Tracks in Mammalian Cells , 2009, Radiation research.

[8]  Hirohiko Tsujii,et al.  Particle radiation therapy using proton and heavier ion beams. , 2007, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[9]  M Scholz,et al.  Biological Imaging of Heavy Charged-Particle Tracks , 2003, Radiation research.

[10]  Ana Pombo,et al.  Chromosome organization: new facts, new models. , 2007, Trends in cell biology.

[11]  F. O. Fackelmayer,et al.  Local Chromatin Mobility is Independent of Transcriptional Activity , 2006, Cell cycle.

[12]  S. Bekker-Jensen,et al.  Dynamic assembly and sustained retention of 53BP1 at the sites of DNA damage are controlled by Mdc1/NFBD1 , 2005, The Journal of cell biology.

[13]  Osamu Onodera,et al.  Aprataxin, a novel protein that protects against genotoxic stress. , 2004, Human molecular genetics.

[14]  J. Aten,et al.  Dynamics of DNA Double-Strand Breaks Revealed by Clustering of Damaged Chromosome Domains , 2004, Science.

[15]  James L. Chen,et al.  Image-Based Modeling Reveals Dynamic Redistribution of DNA Damage into Nuclear Sub-Domains , 2007, PLoS Comput. Biol..

[16]  R K Sachs,et al.  Review: proximity effects in the production of chromosome aberrations by ionizing radiation. , 1997, International journal of radiation biology.

[17]  James G. McNally,et al.  Changes in chromatin structure and mobility in living cells at sites of DNA double-strand breaks , 2006, The Journal of cell biology.

[18]  T. Cremer,et al.  Quantitative motion analysis of subchromosomal foci in living cells using four-dimensional microscopy. , 1999, Biophysical journal.

[19]  K M Prise,et al.  A review of dsb induction data for varying quality radiations. , 1998, International journal of radiation biology.

[20]  David J. Chen,et al.  Repair of HZE-Particle-Induced DNA Double-Strand Breaks in Normal Human Fibroblasts , 2008, Radiation research.

[21]  J. Bartek,et al.  Imaging of protein movement induced by chromosomal breakage: tiny ‘local’ lesions pose great ‘global’ challenges , 2005, Chromosoma.

[22]  Marco Durante,et al.  Cancer risk from exposure to galactic cosmic rays: implications for space exploration by human beings. , 2006, The Lancet. Oncology.

[23]  F. A. Cucinotta,et al.  Immunofluorescence Detection of Clustered γ-H2AX Foci Induced by HZE-Particle Radiation , 2005, Radiation research.

[24]  M. Barcellos-Hoff,et al.  Image-Based Modeling Reveals Dynamic Redistribution of DNA Damage into Nuclear Sub-Domains , 2007, PLoS Comput. Biol..

[25]  J. Savage.  Interchange and intra‐nuclear architecture , 1993, Environmental and molecular mutagenesis.

[26]  S. Kozubek,et al.  Chromatin dynamics during DSB repair. , 2007, Biochimica et biophysica acta.

[27]  Karsten Rippe,et al.  Nuclear body movement is determined by chromatin accessibility and dynamics. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[28]  T. Misteli,et al.  Mobility and immobility of chromatin in transcription and genome stability. , 2007, Current opinion in genetics & development.

[29]  G. Taucher‐Scholz,et al.  Interaction of heavy ions with nuclear chromatin: Spatiotemporal investigations of biological responses in a cellular environment , 2006 .

[30]  M. Nikiforova,et al.  Proximity of chromosomal loci that participate in radiation-induced rearrangements in human cells. , 2000, Science.

[31]  M. Cornforth.  Radiation-Induced Damage and the Formation of Chromosomal Aberrations , 1998 .

[32]  D. Spector,et al.  53BP1 promotes non-homologous end joining of telomeres by increasing chromatin mobility , 2008, Nature.

[33]  K. Mills,et al.  Heavy ion carcinogenesis and human space exploration , 2008 .

[34]  Michael Unser,et al.  A pyramid approach to subpixel registration based on intensity , 1998, IEEE Trans. Image Process..

[35]  D. Alloni,et al.  Heavy-ion effects: from track structure to DNA and chromosome damage , 2008 .

[36]  Burkhard Jakob,et al.  Autophosphorylation of DNA-PKCS regulates its dynamics at DNA double-strand breaks , 2007, The Journal of cell biology.

[37]  C Cremer,et al.  Morphology and dynamics of chromosome territories in living cells. , 2001, Biochimica et biophysica acta.

[38]  A. Murray,et al.  Interphase chromosomes undergo constrained diffusional motion in living cells , 1997, Current Biology.

[39]  N. Gueven,et al.  Live Cell Imaging of Heavy-Ion-Induced Radiation Responses by Beamline Microscopy , 2005, Radiation research.

[40]  R. T. Bree,et al.  The MRN complex: coordinating and mediating the response to broken chromosomes , 2003, EMBO reports.

[41]  M. Lagally,et al.  In situ visualization of DNA double-strand break repair in human fibroblasts. , 1998, Science.

[42]  M. Vázquez,et al.  Chromosomes are predominantly located randomly with respect to each other in interphase human cells , 2002, The Journal of cell biology.

[43]  H. Paretzke,et al.  First steps towards systems radiation biology studies concerned with DNA and chromosome structure within living cells , 2008, Radiation and environmental biophysics.

[44]  H. Tanke,et al.  Chromatin movement visualized with photoactivable GFP-labeled histone H4. , 2008, Differentiation; research in biological diversity.

[45]  M. Cornforth,et al.  Perspectives on the formation of radiation-induced exchange aberrations. , 2006, DNA repair.

[46]  J. Rouse,et al.  Characterisation of the sites of DNA damage-induced 53BP1 phosphorylation catalysed by ATM and ATR. , 2007, DNA repair.

[47]  Wendy A Bickmore,et al.  Chromatin Motion Is Constrained by Association with Nuclear Compartments in Human Cells , 2002, Current Biology.

[48]  M. Durante,et al.  Relationship between radiation-induced aberrations in individual chromosomes and their DNA content: effects of interaction distance , 2001, International journal of radiation biology.